key: cord-0783423-h377ticw
authors: Hong, Kyung Soo; Ahn, June Hong; Jang, Jong Geol; Lee, Jong Ho; Kim, Hong Nam; Kim, Dongha; Lee, Wonhwa
title: GSK-LSD1, an LSD1 inhibitor, quashes SARS-CoV-2-triggered cytokine release syndrome in-vitro
date: 2020-11-17
journal: Signal Transduct Target Ther
DOI: 10.1038/s41392-020-00391-5
sha: b1d47cac9b7bb5b768ae6b4259bc57a583b3f248
doc_id: 783423
cord_uid: h377ticw

nan

the dexamethasone inhibited the expression of pro-inflammatory cytokines by inhibiting the JAK-STAT pathway. 5 Furthermore, our previous studies found that Go6976 , an inhibitor of PKCα that phosphorylates LSD1, and GSK-LSD1, which directly inhibits activation of LSD1, could improve survival rate in the septic mouse model by inhibiting the PKCα-LSD1-NF-κB pathway. 3 In this regard, we treated the PBMCs isolated from severe COVID-19 patients with Go6976/GSK-LSD1. Interestingly, the total level of LSD1 expression remained unchanged, but the phosphorylated-S112 LSD1 was reduced, and thus the LSD1 phosphorylation ratio was also significantly reduced ( Fig. 1a-c) . The Go6976 and GSK-LSD1 also decreased the expression of nuclear NF-κB p65 and ameliorated the cell viability of severe COVID-19 PBMCs (Fig. 1e) . Therefore, we confirmed the Go6976 or GSK-LSD1 as potential therapeutics for severe COVID-19 patients, which can suppress the demethylation of NF-κB p65 by LSD1 and the expression of proinflammatory cytokine genes in vitro. Similar to the results of previous study, 3 in severe COVID-19 PBMCs, as in other infectious diseases, NF-κB p65 is stabilized through demethylation of LSD1, along with increased acetylation of NF-κB p65, an activated form. Whereas, when severe COVID-19 PBMCs were treated with Go6976 and GSK-LSD1, demethylation of NF-κB p65 by LSD1 was suppressed, and methylation was maintained. As a result, the stability of NF-κB p65 was reduced, and the acetylation of NF-κB p65 was also significantly reduced (Fig. 1f) .

Because the drug is currently undergoing phase I clinical assessments an agent for cancer therapy (ClinicalTrials.gov), 6 we intensively evaluated whether GSK-LSD1 has an inhibitory effect on cytokine production. It provides important preclinical information on the shortening of the drug development process through drug repositioning. GSK-LSD1 attenuated the expression of NF-κBdependent pro-inflammatory cytokine genes in severe COVID-19 PBMCs (Fig. 1g ). In addition, as a result of confirming cytokine profiling produced from severe COVID-19 PBMCs (Fig. 1h) , it was verified that the production of cytokines was attenuated by significantly suppressing the activity of NF-κB p65 (Fig. 1i) . After the treatment with GSK-LSD1, quantitative analysis of secreted cytokines demonstrated a markedly decrease in the selectively upregulated levels of pro-inflammatory cytokines such as IL-1β, IL-4, IL-6, MCP-1, IFN-γ, and TNF-α (Fig. 1j-o) .

In previous study, we found that inhibitors of the PKCα-LSD1-NF-κB signaling pathway, 3 Go6976 and GSK-LSD1, that inhibited the expression of several cytokines at once in severe COVID-19 PBMCs. These results may overcome the limitations of existing single cytokine inhibitor for improved suppression of ARDS and sepsis in severe COVID-19 patients. In addition, GSK-LSD1, which has already been undergoing clinical approval as cancer therapy, 6 can be applied directly to severe COVID-19 patients, thereby saving time and cost, and be used as a therapeutic agent that can decrease mortality of severe COVID-19. The best treatment strategy to reduce the COVID-19 severity is to co-administer Fig. 1 GSK-LSD1 inhibits NF-κB p65-mediated cytokine storm in PBMCs isolated from severe COVID-19 patients. PBMCs were isolated from 20 healthy volunteers and 20 severe COVID-19 patients, respectively. a-c Level of total LSD1 (a), phosphorylated-S112 LSD1 (b), and phosphorylation ratio of LSD1 in normal and severe COVID-19 PBMCs (c). d NF-κB p65 activity in normal and severe COVID-19 PBMCs. e Viability of PBMCs isolated from severe COVID-19 patients. f Immunoblot of phosphorylated-S112 LSD1, LSD1, acetylated K314/315-, monomethylated K314/315-NF-κB p65, and nucleus NF-κB p65 was shown in Go6976 or GSK-LSD1-treated PBMCs from normal and severe COVID-19 PBMCs. g Heat map of inflammation-related genes in PBMCs from severe COVID-19 patients after GSK-LSD1 treatment (n = 3). h Cytokines profiling of pharmacological effect of GSK-LSD1 for preventing cytokine storm. i Binding activity of NF-κB p65 in normal and severe COVID-19 PBMCs. j-o Plasma cytokines levels in normal and GSK-LSD1-treated severe COVID-19 PBMCs. Data are reported as mean ± SEM. Significance was set at **P < 0.01 vs. normal; # P < 0.05, ## P < 0.01 vs. severe COVID-19 

COVID-19: consider cytokine storm syndromes and immunosuppression

The pathogenesis and treatment of the 'Cytokine Storm' in COVID-19

PKCalpha-LSD1-NF-kappaB-Signaling cascade is crucial for epigenetic control of the inflammatory response

Dexamethasone in hospitalized patients with COVID-19 -preliminary report

Effect of dexamethasone on NF-kB activation, tumor necrosis factor formation, and glucose dyshomeostasis in septic rats

LSD1/KDM1A inhibitors in clinical trials: advances and prospects

We thank the patients who participated in this study. 

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